1.4 The calcium signaling machinery in N. crassa
1.4.7 Ca 2+ and/or CaM binding proteins
are involved in the control of [Ca2+]c and contain multiple conserved domains, including an N-terminal catalytic domain, a Ca2+ binding EF-hand domain, an autoinhibitory domain, and an overlapping CaM binding domain (Tamuli et al. 2013). CaMK-1, CaMK-2, CaMK-3, and CaMK-4 are four Ca2+/CaM dependent kinases encoded by the NCU09123, NCU02283, NCU06177, and NCU09212 genes, respectively, in N. crassa (Galagan et al. 2003). CaMK-1 mutants have a phase delay, a light-induced phase shift in the circadian conidiation rhythm, a short period lengthening, and a delayed growth phenotype (Yang et al. 2001). The Ca2+/CaM dependent kinases in N. crassa are involved in growth, oxidative stress survival, and other processes (Kumar and Tamuli 2014). Calcineurin is a heterodimer containing a CaM binding catalytic subunit, calcineurin A (CNA-1), and a Ca2+ binding regulatory subunit, calcineurin B (CNB-1), and these two subunits interact physically in N. crassa (Klee et al.
1979; Winkler et al. 1984; Tamuli et al. 2016). Calcineurin is involved in the maintenance of an apical Ca2+ gradient as well as growth and development in N. crassa (Prokisch et al. 1997; Kothe and Free 1998; Tamuli et al. 2016). The NCU04379 gene encodes the Ca2+ and/or CaM binding neuronal calcium sensor-1 (NCS-1), which is involved in growth, Ca2+ stress tolerance, UV survival, and germling fusion (Deka et al. 2011; Palma-Guerrero et al. 2013). The NCU08980 gene encodes another Ca2+ and/or CaM binding protein, NDE-1, which is an external NADPH dehydrogenase in N. crassa mitochondria (Melo et al. 1999, 2001; Borkovich et al. 2004; Carneiro et al. 2007). The NADPH oxidation activity of the NDE-1 protein is Ca2+-dependent and features a Ca2+ binding motif (Melo et al. 1999, 2001).
An Introduction to Neurospora crassa Biology and Calcium Signaling CHAPTER 1
Table 1.1: Ca2+ signaling proteins in N. crassa Sl.
No.
NCU No.
Gene name
Protein name
Protein type Best overallb (e-value;
organism; protein name;
accession number)
1 02762 cch-1 Cch-1 Ca2+ permeable
channel
0; Verticillium dahlia (Cch1);
EGY18507.1
2 06703 mid-1 MID-1 Ca2+ permeable
channel
2e-97; Paracoccidioides brasiliensis (MID-1);
EEH23338.1
3 11680 Ca2+ permeable
channel
0; Ajellomyces dermatitidis (Yvc1); EGE78766.1 4 03305 nca-1 NCA-1 Ca2+-ATPase 0; Trichophyton tonsurans
(SCA-1); EGD96734.1
5 04736 nca-2 NCA-2 Ca2+-ATPase 0; Magnaporthe oryzae (Plasma membrane calcium transporting ATPase 3); EHA56671.1 6 05154 nca-3 NCA-3 Ca2+-ATPase 0; Glomerella graminicola
(Calcium translocating P-type ATPase); EFQ29373.1 7 03292 pmr-1 PMR-1 Ca2+-ATPase 0; Uncinocarpus reesii (PMR-
1); XP_002541437.1
8 08147 ph-7 PH-7 Ca2+-ATPase 0; G. graminicola
(Potassium/sodium efflux P- type ATPase); EFQ36596.1 9 04898 trm-9 TRM-9 Ca2+-ATPase 0; Cordyceps militaris (Cation
transporting ATPase 4);
EGX91104.1 10 03818 trm-10 TRM-10 Ca2+-ATPase 0; V. dahlia (Neo1p);
EGY18069.1
An Introduction to Neurospora crassa Biology and Calcium Signaling CHAPTER 1 11 07966 trm-1 TRM-1 Cation-ATPase 0; T. tonsurans (Cta3p);
EGD97988.1
12 10143 atp-11 ATP-11 Cation-ATPase 0; C. militaris (ATPase type 13A2); EGX92563.1
13 07075 cax CAX Ca2+/H+
exchanger
0; G. graminicola
(Calcium/proton exchanger);
EFQ30300.1 14 00916 trm-15 TRM-15 Ca2+/H+
exchanger
2e-176; Aspergillus fumigatus (Membrane bound cation transporter); XP_001481534.1 15 00795 trm-14 TRM-14 Ca2+/H+
exchanger
1e-149; A. niger (Membrane bound cation transporter);
XP_001400827.2 16 06366 trm-18 TRM-18 Ca2+/H+
exchanger
0; Sclerotinia sclerotiorum (Ca2+/H+ antiporter);
XP_001589752.1 17 07711 trm-19 TRM-19 Ca2+/H+
exchanger
4e-160; T. tonsurans (Vacuolar calcium ion transporter/H+ exchanger); EGD98067.1 18 05360 trm-17 TRM-17 Ca2+/H+
exchanger
0; Metarhizium anisopliase (Calcium permease);
EFY95914.1 19 02826 trm-16 TRM-16 Ca2+/Na+
exchanger
0; V. albo-atrum
(Sodium/calcium exchanger protein); XP_003004985.1 20 08490 trm-20 TRM-20 Ca2+/Na+
exchanger
1e-83; A. niger
(Sodium/calcium transporter);
XP_001397155.1 21 01266 plc-2 PLC-2 Phospholipase C 0; Sordaria macrospora
(Phosphoinositide-specific phospholipase C);
An Introduction to Neurospora crassa Biology and Calcium Signaling CHAPTER 1 22 06245 plc-1 PLC-1 Phospholipase C 0; G. graminicola
(Phosphatidylinositol-specific phospholipase C); EFQ28596.1 23 11415 inl-7 INL-7 Phospholipase C 0; G. graminicola
(Phosphatidylinositol-specific phospholipase C); EFQ31595.1 24 02175 inl-15 INL-15 Phospholipase C 3e-125; Botryotinia fuckeliana
(BcPLC2); CCD34776.1
25 04120 cmd CaM Calmodulin 1e-103; Gibberella zeae (CaM);
XP_382067.1 26 03804 cna-1 CNA-1 Ca2+ and/or CaM
binding protein
0; S. macrospora
(Serine/threonine-protein phosphatase 2B catalytic subunit protein);
XP_003352213.1 27 03833 cnb-1 CNB-1 Ca2+ and/or CaM
binding protein
2e-119; Trichoderma reesei (Calcineurin beta subunit);
EGR44907.1 28 09265 cnx-1 CNX-1 Ca2+ and/or CaM
binding protein
0; S. macrospora (cnx1);
XP_003347545.1 29 05225c nde-1 NDE-1 Ca2+ and/or CaM
binding protein
0; M. oryzae (Mitochondrial NADH dehydrogenase);
EHA47323.1
30 02115 Ca2+ and/or CaM
binding protein
0; M. oryzae (EF-hand domain containing protein);
EHA48778.1 31 01564 mic-4 MIC-4 Ca2+ and/or CaM
binding protein
0; M. oryzae (Calcium
dependent mitochondrial carrier protein); EHA48778.1
32 06948 Ca2+ and/or CaM
binding protein
2e-54; Mycosphaerella graminicola (Calcium ion
An Introduction to Neurospora crassa Biology and Calcium Signaling CHAPTER 1 binding, calmodulin);
EGP88834.1) 33 04379 ncs-1 NCS-1 Ca2+ and/or CaM
binding protein
4e-126; Grosmannia clavigera (Neuronal calcium sensor 1);
EFX03580.1 34 02738 pef-1 PEF-1 Ca2+ and/or CaM
binding protein
2e-130; V. dahlia (Peflin);
EGY21808.1 35 09871 nup-34 NUP-34 Ca2+ and/or CaM
binding protein
4e-33; V. dahlia (Centrin-3);
EGY16271.1 36 01241 mic-2 MIC-2 Ca2+ and/or CaM
binding protein
0; T. reesei (Mitochondrial carrier protein); EGR44893.1 37 06347 ask-2 ASK-2 Ca2+ and/or CaM
binding protein
0; S. macrospora (Actin cytoskeleton–regulatory complex protein);
XP_003350109.1 38 06617 cdc4-2 CDC4-2 Ca2+ and/or CaM
binding protein
7e-93; V. albo-atrum (Myosin regulatory light chain cdc4);
XP_003009631.1 39 03750 cmd-2 CMD-2 Ca2+ and/or CaM
binding protein
8e-74; B. fuckeliana (Calmodulin);
XP_001560827.1 40 08980 nde-2 NDE-2 Ca2+ and/or CaM
binding protein
0; G. clavigera (Alternative NADH-dehydrogenase);
EFX03867.1 41 02283 camk-2 CaMK-2 Ca2+ and/or CaM
binding protein
0; S. macrospora (Calcium/calmodulin-
dependent protein kinase type I); XP_003344498.1
42 09123 camk-1 CaMK-1 Ca2+ and/or CaM binding protein
0; Sporothrix schenckii (Calcium/calmodulin- dependent kinase);
An Introduction to Neurospora crassa Biology and Calcium Signaling CHAPTER 1 43 09212 camk-4 CaMK-4 Ca2+ and/or CaM
binding protein
0; V. dahliae (Serine/threonine- protein kinase srk1);
EGY15110.1 44 02814 prd-4 PRD-4 Ca2+ and/or CaM
binding protein
0; G. clavigera
(Serine/threonine-protein kinase chk2); EFX01629.1 45 06650 spp-3 SPP-3 Ca2+ and/or CaM
binding protein
3e-61; Nectria haematococca (Phospholipase A2);
XP_003042542.1
46 02411 Ca2+ and/or CaM
binding protein
0; G. graminicola (Microtubule-associated protein); EFQ31793.1 47 06177 camk-3 CaMK-3 Ca2+ and/or CaM
binding protein
0; M. grisea (CMKK2);
ACM41720.1
48 04265 inv INV Ca2+ and/or CaM
binding protein
7e-85; Bacillus megaterium (Betafructosidase FruA);
AEN90524.1
aAdapted from Tamuli et al. 2013. bBLASTp search was performed at NCBI (http://blast.ncbi.nlm.nih.gov/Blast.cgi; Altschul et al. 1990, 1997, 2005) with default parameters for each of the 48 Ca2+ signaling proteins against the non-redundant protein sequence databases, and the respective best overall match in other organisms with an e-value has been indicated. cNCU05225 was indicated as NCU08980.1 in Borkovich et al. 2004.